Antibiotic resistance, plasmid profile and RAPD-PCR analysis of enteropathogenic Escherichia coli (EPEC) clinical isolates (original) (raw)
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Journal of Medical Microbiology, 2013
Diarrhoea continues to be one of the most common causes of morbidity and mortality among infants and children in developing countries. To investigate the incidence, antimicrobial resistance and genetic relationships of enteropathogenic Escherichia coli (EPEC) in children with diarrhoea, a total of 612 stool specimens were collected in Tehran, Iran, and cultured to isolate strains of EPEC. The disc diffusion method was used to determine the susceptibility of the isolates according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. The presence of eae, stx and bfp-A genes was determined by PCR. The genetic relationships between EPEC isolates were determined by pulsed-field gel electrophoresis (PFGE). Out of the 412 strains of E. coli obtained from 612 diarrhoeal stool specimens, 23 (5.6 %) were identified as EPEC, of which seven (30.4 %) were classified as typical strains of EPEC and 16 (69.6 %) were classified as atypical. Out of the 23 EPEC isolates, 69.5 % were resistant to ampicillin, 39.1 % were resistant to tetracycline and cotrimoxazole, 30.4 % were resistant to cefpodoxime, ceftazidime, ceftriaxone and aztreonam, and 26.1 % were resistant to imipenem. The isolates were classified into 21 pulsotypes by PFGE profiles. The present study shows that typical and atypical EPEC isolates displayed considerable heterogeneity in PFGE profiles and EPEC infections were only sporadic in Tehran. Overall 69 % of isolates were resistant to at least one of the antibiotics tested.
International Journal of Current Microbiology and Applied Sciences
Diarrheal diseases are a major cause of morbidity and mortality in developing countries. In the past decades several new enteric pathogens, including bacterial, viral, and parasitic agents have been described. Escherichia coli (Esch. coli) is one of the leading cause of acute diarrhea in developing countries in children under 5 years old, with significant morbidity and mortality. The present study was carried out in the Department of Microbiology Mayo Institute of Medical Sciences and Research Lucknow. The study group included 81 children suffering from diarrhoea attending the inpatient or outpatient sections of the Department of Paediatrics. Among the bacterial pathogens detected, Esch. coli was the commonest. In the present study major symptoms reported were diarrhea, fever and vomiting. Examination of stool samples showed that 32 (39.5%) children harbored one or more of the Escherichia coli pathotypes. Out of those 32 who were subjected to serotyping 9 (28.12%) were untypable and 5 (15.62%) were rough strains. Of the typable strains, Enteropathogenic E.coli (EPEC) accounted for 3.12%, Enterotoxicogenic E.coli (ETEC) accounted for 12.5%, Enterohaemorragic (EHEC) accounted for 3.12 %. Of the ETEC strains, serogroups O173 (1), O11 (1), O15 (1) and O8 (1) were found. The EPEC strain found was O86 and EHEC was serogroup O172. Among the other serogroups which comprised of 37.5% strains, O161 (3) was predominant. Antibiotic resistance was documented for Ampicillin (100%).
BioMed Research International, 2020
Background. Diarrhea has been reported as the leading cause of childhood mortality and morbidity globally but with disproportionate impacts in developing nations. Among bacterial etiologic agents of diarrhea, diarrheagenic Escherichia coli is the main cause of the disease among children under the age of 5 years. This study is aimed at determining the prevalence and antibiogram pattern of diarrheagenic Escherichia coli (DEC) pathotypes associated with diarrhea cases in the study area. Methods. A total of 120 presumptive isolates of E. coli were obtained from diarrheal stool samples from male and female patients below 12 years of age using chromogenic agar. Confirmation of the isolates and screening for virulence genes were determined by polymerase chain reaction (PCR) while antimicrobial susceptibility testing was performed using the disk diffusion method. The presence of antibiotic resistance genes to chloramphenicol and tetracycline among the confirmed isolates was also profiled by PCR based on the observed phenotypic resistance pattern. Results. Of the 120 presumptive isolates, 88.3% (106/120) were confirmed as E. coli through PCR. The molecular pathotyping of the confirmed isolates showed their distribution as 41% (43/106) of diffusely adhering E. coli (DAEC), 17% (18/106) of enterohemorrhagic E. coli (EHEC), 17% (18/106) of enteropathogenic E. coli (EPEC), and 10% (11/106) of enteroinvasive E. coli (EIEC), while enteroaggregative E. coli (EAEC) and enterotoxigenic E. coli (ETEC) were not detected, and the remaining 15% did not belong to any pathotype. Notably, high resistance of the isolates to commonly used antimicrobials was observed as follows: ampicillin (98%), chloramphenicol (94%), trimethoprimsulfamethoxazole (96%), and tetracycline (90.6%), while a relatively low number of the confirmed isolates were resistant to ciprofloxacin (45%) and imipenem (36%). In addition, 94% of the isolates that exhibited phenotypic resistance against chloramphenicol harbored the catA1 resistance gene while 89% that showed resistance to tetracycline had tetA genes. Conclusions. These findings showed that DEC could be considered as the leading etiologic bacterial agent responsible for diarrhea in the study community, and the observable high degree of resistance of the isolates to antimicrobial agents is of huge significance, calling for stakeholders to adopt and consolidate the existing antimicrobial stewardship scheme of the government, in order to ensure an uncompromised public health.
Iranian Journal of Microbiology, 2019
Background and Objectives: Escherichia coli is a common enteric pathogen of human and livevestock. Antibiotic resistance is the main concern of public health. The aim of this study was to detect this bacterium in stool samples of diarrheal patients and identify the phenotypic and genotypic characterizations of antibiotic-resistant isolates such as dfrA1, sul1, citm, tetA, qnr, aac(3)-IV in Shahrekord. Materials and Methods: Two hundred fifty diarrheal stool samples from patients were collected. Microbiological and biochemical examinations were done to detect E. coli. Phenotypic and genotypic antibiotic resistance of the isolates were determined using dick diffusion method and polymerase chain reaction (PCR), respectively. Results: Among 114 E. coli isolates, the least resistance was for gentamicin (0%) and the most resistance was for trimethoprim (79.8%). The resistance to sulfamethoxazole, ciprofloxacin, ampicillin, and tetracycline were 71.05%, 10.5%, 52.63%, and 3.5% respectively...
Journal of Tropical Medicine, 2022
Introduction. Pathogenic Escherichia coli strains cause diarrheal infection in children due to their virulence factors. A nonanalytical observational study followed by a purposive sampling technique was conducted from October 2017 to June 2018, to determine the antimicrobial susceptibility patterns and molecularly detect pathogenic Escherichia coli strains in under-ve children at Wolaita Sodo town using molecular and the Kirby-Bauer disc di usion method. Result. In the current investigation, out of 110 stool samples, Escherichia coli was isolated in 68 (61.8%) (95% CI: 52.1-70.9%). Out of 68 Escherichia coli isolates, 61.9% of Escherichia coli isolates were resistant, 9.4% were intermediately resistant, and 28.7% were susceptible. Among the antimicrobial agents, 91.2% of Escherichia coli isolates were highly sensitive to cipro oxacin followed by nor oxacin (86.8%). Virulence genes were detected in 55.9% (38/68) (95% CI 52.1%-70.9%) of isolates. e following genes were detected: stx1 (6 (8.8%)), stx2 (4 (5.9%)), eaeA (5 (7.4%)), eaat (14 (20.5%)), and St (4 (5.9%)). Out of 68 Escherichia coli isolates, 43 (63.2%) isolates had shown multidrug resistance patterns. Furthermore, 11.8% of Escherichia coli isolates had shown resistance to eight di erent drugs. e multidrug resistance index value of Diarrheagenic Escherichia coli pathotypes was greater than or equal to 0.4, which indicates the high risk of resistance. Conclusion. is study demonstrated important pathogenic Escherichia coli strains and multidrug resistance in isolates containing virulence genes. Wise use of antimicrobials and improving the hygienic practices amongst parents of children reduce its occurrence. erefore, appropriate usage of antimicrobial agents should also be highly practiced in hospitals.
Jundishapur Journal of Microbiology
Background: Enteropathogenic Escherichia coli (EPEC) is one of the most important pathogens among young children worldwide. Both eae and bfp genes have been used to identify EPEC strains and categorize them into typical and atypical strains. They may be an emerging pathogen in both developing and developed countries. Objectives: This study was primarily conducted to assess the epidemiology, drug resistance, and β-lactamase distribution of EPEC, as well as the detection of efa1/lifA in atypical strains. Methods: A total of 251 E. coli strains isolated from children with diarrhea were evaluated for their EPEC pathotype by PCR for the presence of eae, stx1, stx2, and bfp genes. Serogrouping with polyvalent antisera was performed to confirm EPEC strains. Atypical EPEC-containing samples were evaluated for the efa1/lifA gene. EPEC isolates were assessed to recognize the antibiotic resistance and screened to detect extended-spectrum β-lactamases (ESBLs). Results: Enteropathogenic E. coli ...
Journal of health, population, and nutrition, 2018
Emergence of atypical enteropathogenic Escherichia coli (EPEC) and hybrid E. coli (harboring genes of more than one DEC pathotypes) strains have complicated the issue of growing antibiotic resistance in diarrhoeagenic Escherichia coli (DEC). This ongoing evolution occurs in nature predominantly via horizontal gene transfers involving the mobile genetic elements like integrons notably class 1 integron. This study was undertaken to determine the virulence pattern and antibiotic resistance among the circulating DEC strains in a tertiary care center in south of India. Diarrhoeal stool specimens were obtained from 120 children (< 5 years) and 100 adults (> 18 years), subjected to culture and isolation of diarrhoeal pathogens. Conventional PCR was performed to detect 10 virulence and 27 antimicrobial resistance (AMR) genes among the E. coli isolated. DEC infection was observed in 45 (37.5%) children and 18 (18%) adults, among which [18 (40%), 10 (10%)] atypical EPEC was most commonl...
Jundishapur Journal of Microbiology, 2015
Background: Several studies performed in developed and developing countries have identified enteroaggregative Escherichia coli (EAEC) as the emerging cause of pediatric diarrhea. Objectives: This study investigated the phenotypic and genetic characteristics of EAEC strains isolated from children with diarrhea between 2007-2008 in Tehran, Iran. Materials and Methods: EAEC strains were examined for virulence plasmid genes (aap, aggR, and aatA), biofilm formation, and drug resistance. In addition, pulsed-field gel electrophoresis (PFGE) profiles of these strains were determined. Results: Significant percentage of local EAEC strains carried the virulence plasmid genes and formed biofilms. In addition, these strains showed high resistance to ampicillin (100%), tetracycline (65.7%), streptomycin (58.7%), chloramphenicol (52.6%), and trimethoprim/ sulfamethoxazole (51.7%) and had different PFGE patterns. Conclusions: These results indicated that EAEC strains isolated from Iranian children with diarrhea were heterogeneous and showed high resistance rates against commonly used antibiotics, which was similar to that reported in studies performed in other countries.
International Journal of One Health, 2020
Aim: This study was aimed to determine the virulent genes and antibiotic resistance patterns among circulating diarrheagenic Escherichia coli (DEC) pathotypes in a tertiary care health center in east of Nigeria. Materials and Methods: Diarrheal stool samples were obtained from 80 children under 5 years and E. coli was isolated and identified using standard biochemical and molecular methods. Multiplex polymerase chain reaction (PCR) was used to detect eight virulent genes of DEC. Disk diffusion method was used to determine the antibiotic susceptibility of DEC. Results: DEC infection was observed in 54 (68%) children among which ial gene for enteroinvasive E. coli (EIEC) (40% [n=22]) was commonly detected followed by eltA/eltB for enterotoxigenic E. coli (ETEC) (30% [n=16]), pCVD for enteroaggregative E. coli (EAEC) (20% [n=11]), and eaeA/bfpA for typical enteropathogenic E. coli (EPEC) (10% [n=5]). The DEC isolates phenotypically exhibited resistance for ampicillin (AMP) (44 [81%]), followed by ciprofloxacin (CIP)/ levofloxacin (LEV) (28 [52%]), cefoxitin (FOX) (11 [20%]), and amoxicillin-clavulanic acid (AMC) (6 [11%]). About 60% isolates of stable toxins-ETEC were resistant to AMC, CIP, and LEV while all the labile toxin-ETEC exhibited resistance to AMP. About 60% (n=6) resistance were seen in EAEC against ampicillin, AMC, FOX, CIP, and LEV. In EIEC, all the isolates (n=22) were resistant to AMP while 50% (n=11) were resistant to both CIP and LEV. All EPEC (n=5) were resistant to AMP, FOX, CIP, and LEV. Conclusion: High frequency of virulent ial and eltA/eltB genes for EIEC and ETEC, respectively, suggests that they are the primary etiological agents of diarrhea in children among DEC pathotypes. Resistance of DEC to more than two classes of antibiotics indicate possible emergence of multidrug resistance.
Ibrahim Medical College Journal, 2015
Enteropathogenic Escherichia coli (EPEC) mediated infantile diarrhoea among children is an important cause of morbidity and mortality in developing countries. The antimicrobial susceptibility pattern of EPEC strains isolated from children under 5 years of age was studied. Stool samples from 272 patients with diarrhoea were collected from two tertiary care hospitals. Out of 272 stool samples, 20 (7.35%) isolates were identified as EPEC on the basis of presence of bfpA gene detected by polymerase chain reaction and antibiotic susceptibility testing was performed on these EPEC strains by Kirby-Bauer disc diffusion method. The antimicrobial susceptibility test revealed that the EPEC isolates were highly resistant to ampicillin (100%), nalidixic acid (95%) and tetracycline (95%) and were sensitive to ceftazidime (95%), cefotaxime (90%), ceftriaxone (95%), imipenem (100%) and levofloxacin (85%). Isolation of EPEC is of great importance since they are responsible for acute diarrhoeal diseases in large number of children under the age of five years. The high antimicrobial resistance observed in our study indicates indiscriminate or improper use of antimicrobials, besides the risks of self-medication.